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1.
Indian J Microbiol ; 62(1): 61-69, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-35068605

RESUMEN

Deoxynivalenol (DON) is synthesized by Fusarium species that frequently infect crops during storage, and it's harm risk to human is reflected in the consumption of infected food crops or indirectly through foods of animal origin. In this study, Hela and Chang liver cells were used to research the cellular apoptosis induced by deoxynivalenol. Cells were treated by DON toxin with a series of concentration and incubated for different time. MTT, fluorescence microscope, flow cytometer and Western blot methods were used to analyze the effect of DON on the cell apoptosis in vitro and in vivo systematically. The results showed that DON was toxic to the cells tested. After being treated by DON, the morphology of Chang livers and Hela cells changed significantly. The DON promoted apoptosis in a dose- and time-dependent manner. The activity of Caspase 3 was significantly increased in DON-induced apoptosis. Moreover, endogenous Glutathione (GSH) level in these cell lines was gradually decreased. In the early apoptosis progress, oxidative stress was induced by DON. When DON reached 10 µg/mL, a markedly increased content of Malondialdehyde (MDA) was detected in both Hela and Chang liver cells. Furthermore, an in vivo test indicated that DON had toxicity to mice by causing weight loss and swollen spleen, and significantly increased expression of AST and ALT. In conclusion, the DON was toxic to mice and could induce the apoptosis of tested cells undergoing a Caspase-3 related pathway.

2.
ACS Omega ; 4(12): 15112-15119, 2019 Sep 17.
Artículo en Inglés | MEDLINE | ID: mdl-31552356

RESUMEN

Compared to inorganic mercury (Hg2+), methyl-mercury (CH3Hg+) and ethyl-mercury (C2H5Hg+) (organic mercury) not only have a much stronger toxicity but also are more easily accumulated by marine organisms to produce bioamplification. Therefore, the simultaneously onsite detection of Hg2+ and organic mercury is of great significance to ensure the safety of seafood, and it is also a hard challenge. We designed a T-rich aptamer, HT7, for specifically recognizing Hg2+ and organic mercury and developed a multicolor aptasensor for simultaneous discrimination and detection of Hg2+ and organic mercury with only bare-eye observation using HT7 as a recognition probe and gold nanorods (AuNRs) as a signal. In the presence of Hg2+ and Ag+, Hg2+ preferentially and specifically bind with HT7 immobilized on AuNRs surface and induce the formation of a monolayer Ag/Hg amalgam on the AuNRs surface after reduction, resulting in a change in color from orange to faint purple and a corresponding shift in the absorption peak from 820 to 730 nm in the solution. However, in the presence of CH3Hg+ or C2H5Hg+ and Ag+, CH3Hg+ or C2H5Hg+ preferentially bind with HT7 immobilized on the AuNRs surface and induce the formation of a monolayer Ag0 on the AuNRs surface after reduction, which results in the change in color from orange to atrovirens and the corresponding shift in the absorption peak shift from 820 to 670 nm in the solution. Thus, the inorganic and organic mercury (total of CH3Hg+ and C2H5Hg+) can be specifically discriminated and detected by only bare-eye observation. The method can be used to simultaneously detect inorganic and organic mercury in seawater by the bare-eye observation with a visual detection limit of 2.0 ppm for Hg2+ and 10.0 ppm for organic mercury. The success of this study is a useful enlightenment to develop an instrument-free method for an onsite detection of trace inorganic and organic mercury in environment by a bare-eye observation, although the sensitivity of the method is relatively low.

3.
Toxicon ; 108: 210-5, 2015 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-26525659

RESUMEN

Fumonisin B1 (FB1) is the most common and highest toxic of fumonisins species, exists frequently in corn and corn-based foods, leading to several animal and human diseases. Furthermore, FB1 was reported that it was associated with the human esophageal cancer. In view of the harmful of FB1, it is urgent to develop a feasible and accuracy method for rapid detection of FB1. In this study, a competitive immunoassay for FB1 detection was developed based on colloidal gold-antibody conjugate. The FB1-keyhole limpet hemoeyanin (FB1-KLH) conjugate was embedded in the test line, and goat anti-mouse IgG antibody embedded in the control line. The color density of the test line correlated with the concentration of FB1 in the range from 2.5 to 10 ng/mL, and the visual limit detection of test for FB1 was 2.5 ng/mL. The results indicated that the test strip is specific for FB1, and no cross-reactivity to other toxins. The quantitative detection for FB1 was simple, only needing one step without complicated assay performance and expensive equipment, and the total time of visual evaluation was less than 5 min. Hence, the developed colloidal gold-antibody assay can be used as a feasible method for FB1 rapid and quantitative detection in corn samples.


Asunto(s)
Monitoreo del Ambiente/métodos , Fumonisinas/análisis , Fusarium/metabolismo , Oro Coloide/química , Zea mays/microbiología , Monitoreo del Ambiente/instrumentación , Fumonisinas/química , Fumonisinas/aislamiento & purificación , Inmunoensayo/instrumentación , Inmunoensayo/métodos
4.
Talanta ; 138: 15-19, 2015 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-25863365

RESUMEN

An enzyme-free and label-free fluorescent biosensor is developed by G-quadruplex-based hybridization chain reaction (HCR) for small molecules, using adenosine triphosphate (ATP) as the model. Aptamer probes for the recognition of small molecules are hybridized with blocking probes. The G-quadruplex sequences are incorporated into one of the two HCR hairpin probes. In the presence of small molecules (ATP), the formation of aptamer-ATP bioaffinity complexes induces the release of blocking probes; the released blocking probes initiate HCR and numerous G-quadruplexes along DNA nanowires are self-assembled after the HCR process. Using N-methyl mesoporphyrin IX (NMM) as the fluorophore, a "turn-on" fluorescence response can be achieved and detected as low as 15 µmol L(-1) of ATP. This biosensor is applied to detect ATP in biologic samples with satisfactory results.


Asunto(s)
Adenosina Trifosfato/orina , Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , Colorantes Fluorescentes/química , G-Cuádruplex , Nanocables/química , Hibridación de Ácido Nucleico/métodos , Fluorescencia , Humanos , Mesoporfirinas/química , Espectrometría de Fluorescencia/métodos
5.
Biosens Bioelectron ; 71: 256-260, 2015 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-25913446

RESUMEN

A monoclonal hybridoma cell named 5B9 against tetrodotoxin (TTX) was obtained after fusion of myeloma SP2/0 cells with spleen cells isolated from the immunized Balb/c mice. The 5B9 monoclonal antibody (McAb) with high affinity (about 2.55 × 10(9)) is specific to TTX, and this McAb belongs to the immunoglobulin G (IgG) isotype. Finally, an enzyme-linked immunosorbent assay (ELISA) and colloidal gold immunoassay were established based on this McAb. The linear range of ELISA to detect TTX was 5-500 ng/mL, and the limit of detection (LOD) was 4.44 ng/mL. The average CV of intra- and inter-assay was less than 8%, with the samples recovery range of 70.93-99.99%. A competitive format colloidal gold strip was developed for detection of TTX in real samples, and the LOD for TTX is 20 ng/mL, and the assay time of the qualitative test can be finished in less than 10 min without any equipment. The result from test strip revealed that the test strip has a good agreement with those obtained from ELISA.


Asunto(s)
Anticuerpos Monoclonales/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Oro Coloide/química , Toxinas Marinas/análisis , Tetrodotoxina/análisis , Animales , Anticuerpos Monoclonales/química , Femenino , Hibridomas/inmunología , Límite de Detección , Toxinas Marinas/inmunología , Ratones Endogámicos BALB C , Tetraodontiformes , Tetrodotoxina/inmunología
6.
Artículo en Inglés | MEDLINE | ID: mdl-22119123

RESUMEN

In this study, the noncovalent functionalization of graphene with cationic porphyrins in an aqueous medium was investigated using UV-vis and fluorescence approaches. To characterize the interaction between graphene and cationic porphyrins, 5,10,15,20-tetra (4-pyridyl)-21H,23H-porphine, 5,10,15,20-tetrakis (1-methyl-4-pyridinio) porphyrin tetra (p-toluenesulfonate) and 5,10,15,20-tetrakis (4-trimethylammoniophenyl) porphyrin tetra (p-toluenesulfonate) porphyrin were chosen as reagents. The intermolecular interactions were found to occur immediately after mixing the cationic porphyrins with graphene. The absorption spectra of the cationic porphyrins after mixing with graphene showed distinct red shifts of the Soret and Q-bands compared to free cationic porphyrins indicating that interactions occur between the cationic porphyrins and graphene. A strong fluorescence quenching of the cationic porphyrins in the presence of graphene indicated that efficient electron or energy transfer occurred from the excited state of the cationic porphyrins to graphene. Cationic porphyrins were immobilized on the surface of graphene through electrostatic and π-π stacking interactions, and the chemical shape of graphene played an important role in the intermolecular interactions and the red shift extent of cationic porphyrins is mostly dependent on the functional groups and charges of the graphene surface. The results show that less functional groups on the graphene's surface and edge would lead to stronger π-π stacking interactions between graphene and cationic porphyrins.


Asunto(s)
Grafito/química , Porfirinas/química , Absorción , Ácidos Carboxílicos/química , Cationes , Microscopía de Fuerza Atómica , Espectrometría de Fluorescencia
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